Harada Koji, Kawaguchi Shin-Ichi, Onoue Tomitaro, Kawashima Yuichiro, Yoshida Hideo, Sato Mitsunobu
Second Department of Oral and Maxillofacial Surgery, University of Tokushima, School of Dentistry, 3-18-15 Kuramoto-cho, Tokushima 770-8504, Japan.
Oncol Rep. 2005 Feb;13(2):201-6.
In this study, we attempted to use a non-viral gene transfer system, in vivo electroporation, in oral cancer cell B88 xenografts. To evaluate this in vivo gene transfer method, the GFP gene was transfected into xenografts by electroporation. Then, the efficiency of transfection of exogenous p27Kip1 gene by electroporation was confirmed by Western blot analysis. Next, to estimate the reduction of oral cancer xenografts by this method, we measured the size of B88 xenografts in nude mice after electroporation with the wild- or mutant-type p27Kip1 gene. The growth of tumors was markedly suppressed by mutant-type p27Kip1 gene transfection by electroporation compared with transfection of wild-type p27Kip1 gene or empty vector only. Moreover, histological specimens revealed apoptotic cell death was increased in mutant-type p27Kip1-transfected tumors compared to wild-type or empty vector only. These results suggest that it is possible to transfer mutant-type p27Kip1 into oral cancer xenografts using electroporation and to suppress the growth of tumors, furthermore, it is suggested that this system might be used for oral cancer.
在本研究中,我们尝试在口腔癌细胞B88异种移植瘤中使用一种非病毒基因转移系统——体内电穿孔法。为评估这种体内基因转移方法,通过电穿孔将绿色荧光蛋白(GFP)基因转染至异种移植瘤中。然后,通过蛋白质免疫印迹分析证实了通过电穿孔转染外源性p27Kip1基因的效率。接下来,为评估该方法对口腔癌异种移植瘤的抑制作用,我们在用野生型或突变型p27Kip1基因进行电穿孔后,测量了裸鼠体内B88异种移植瘤的大小。与仅转染野生型p27Kip1基因或空载体相比,通过电穿孔转染突变型p27Kip1基因可显著抑制肿瘤生长。此外,组织学标本显示,与仅转染野生型或空载体相比,转染突变型p27Kip1基因的肿瘤中凋亡细胞死亡增加。这些结果表明,利用电穿孔法将突变型p27Kip1基因导入口腔癌异种移植瘤并抑制肿瘤生长是可行的,此外,该系统可能适用于口腔癌。